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C Oliver

Publications and source records attributed to C Oliver.

At least 271 records · Page 15Linked to original sources

Interaction of opiates with opioid binding sites in the bovine adrenal medulla: II. Interaction with kappa sites.

In this study we examined the interaction of opiates with kappa binding sites in the bovine adrenal medulla. [3H]Ethylketocyclazocine (EKC), [3H]etorphine, and [3H]bremazocine stereoselective bindings were used to assay these interactions. The kappa sites were found to be heterogeneous: [3H]bremazocine identified with high affinity all subtypes of these sites. [3H]EKC, in the presence of saturating concentrations of [D-Ala2, D-Leu5]-enkephalin (DADLE) (5 microM), was used to identify kappa 1 sites, on which dynorphin A (1-13) bound with high affinity. Either [3H]EKC or [3H]etorphine in the presence of 5 microM DADLE identified the kappa 2 subtype. This subtype was found to interact with beta-endorphin and especially with the octapeptide Met5-enkephalyl-Arg6-Gly7-Leu8. Furthermore, [3H]etorphine identified in the bovine adrenal medulla a third high-affinity component, in the presence of 5 microM DADLE. This residual interaction was found to be equally stereoselective and presenting kappa selectivity. Met5-enkephalyl-Arg6-Phe7 interacted preferentially with this site. The three kappa subtypes interacted differentially with monovalent (Na+, K+, and Li+) and divalent (Ca2+, Mg2+, and Mn2+) ions by modification of the apparent concentration of the accessible sites and/or by changes of the apparent KD for radioligands. Modifying agents (proteolytic enzymes, thiol-modifying reagents, and A2-phospholipase) produced different effects on each subtype of the kappa site, suggesting a different protein (or protein-lipid?) composition.

Adrenal Medulla↗

[Passive immunization with an anti-oCRF41 immune serum inhibits the circadian increase of plasma ACTH in rats].

For 24 hrs. after i.v. injection of 1 ml of an undiluted immune serum raised against oCRF41, the diurnal surge of plasma ACTH dropped to a short-lived limited rise above baseline level. On the second day after injection, the ACTH level in treated rats rose to a subnormal level, although both plasma dilution of the immune serum and its binding capacity in the plasma remained unchanged throughout the experiment. Plasma corticosterone, on the contrary, displayed a normal circadian rhythm during the entire experiment. However, in animals given a second injection of 0.5 ml oCRF41 immune serum 32 hrs. after the first, both ACTH and corticosterone titers fell rapidly below their circadian minimal levels in controls. Concomitantly, the concentration of immune serum in the peripheral plasma, and its capacity to bind to oCRF, rose by 50%. The major role of CRF41 as a diurnal trigger of the circadian rhythm of ACTH is discussed, as well as the limits of passive immunization.

Adrenocorticotropic Hormone↗

Acute megakaryoblastic leukemia in childhood.

Routine morphologic and cytochemical study of the leukemic cells of a 13-month-old child did not permit definitive determination of either a lymphoid or a myeloid cell origin. However, ultrastructural cytochemical analysis revealed platelet peroxidase (PPO) reactivity. The malignant cells expressed PPO in both unfixed and fixed preparations. PPO was observed in the perinuclear space and reticulum but not in the Golgi saccules. In a 1% glutaraldehyde, 1% formaldehyde solution known to inhibit PPO but not myeloperoxidase, no reaction product was observed. The demonstration of PPO activity in these undifferentiated cells established their megakaryoblastic lineage. Further studies including DNA flow cytometry confirmed that these cells were megakaryoblasts. This study demonstrates that megakaryoblastic malignancy may occur as an acute leukemia of childhood and emphasizes the value of ultrastructural cytochemistry in cases of acute leukemia which defy routine classification. Abnormalities of chromosome 21 were present in our patient as in four previous cases of megakaryoblastic leukemia in childhood. The nonrandomness of chromosome 21 involvement in this disease should therefore be considered.

Acute Disease↗

Evidence for both prelysosomal and lysosomal intermediates in endocytic pathways.

Horseradish peroxidase (HRP), an enzyme internalized by fluid phase pinocytosis, has been used to study the process by which pinosome contents are delivered to lysosomes in Chinese hamster ovary cells. Pinosome contents were labeled by allowing cells to internalize HRP for 3-5 min. Following various chase times, cells were either processed for HRP and acid phosphatase (AcPase) cytochemistry or homogenized and fractionated in Percoll gradients. In Percoll gradients, pinosomes labeled by a 3-5 min HRP pulse behaved as a vesicle population more dense than plasma membrane and less dense than lysosomes. In pulse-chase experiments, internalized HRP was chased rapidly (3-6 min chase) to a density position intermediate between the "initial" pinocytic vesicle population and lysosomes. With longer chase periods, a progressive accumulation of HRP in more dense vesicles was observed. Correspondence between the HRP distribution and lysosomal marker distribution was reached after a approximately 1-h chase. By electron microscope cytochemistry of intact cells, the predominant class of HRP-positive vesicles after pulse uptakes or a 3-min chase period was characterized by a peripheral rim of reaction product and was AcPase negative. After 10-120-min chase periods, the predominant class of HRP-positive vesicles was characterized by luminal deposits and HRP activity was frequently observed in multivesicular bodies. HRP-positive vesicles after a 10- or 30-min chase were AcPase-positive. No HRP activity was detected in Golgi apparatus. Together these observations indicate that progressive processing of vesicular components of the vacuolar apparatus occurs at both a prelysosomal and lysosomal stage.

Animals↗

A single mutation in Chinese hamster ovary cells impairs both Golgi and endosomal functions.

A Chinese hamster ovary cell mutant DTG 1-5-4, was selected for pleiotropic defects in receptor-mediated endocytosis by methods previously described (Robbins, A. R., S. S. Peng, and J. L. Marshall, 1983, J. Cell Biol., 96:1064-1071). DTG 1-5-4 exhibited increased resistance to modeccin, Pseudomonas toxin, diphtheria toxin, Sindbis virus, and vesicular stomatitis virus, as well as decreased uptake via the mannose 6-phosphate receptor. Fluorescein-dextran-labeled endosomes isolated from DTG 1-5-4 were deficient in ATP-dependent acidification in vitro. Endocytosis and endosome acidification were both restored in revertants of DTG 1-5-4 and in hybrids of DTG 1-5-4 with DTF 1-5-1, another endocytosis mutant exhibiting decreased ATP-dependent endosome acidification. Both DTG 1-5-4 and DTF 1-5-1 were blocked at two stages of infection with Sindbis virus: at low multiplicities of infecting virus, resistance reflected a block in viral penetration into the cytoplasm, but at higher multiplicities of infection the block was in virus release. Like endocytosis, release of Sindbis virus was increased in revertants of DTG 1-5-4 and in DTG 1-5-4 X DTF 1-5-1 hybrids. Decreased release of virus from DTG 1-5-4 correlated with defects in some of the Golgi apparatus-associated steps of Sindbis glycoprotein maturation: proteolytic processing of the precursor pE2, galactosylation, and transport to the cell surface all were inhibited. In contrast, mannosylation, fucosylation, and acylation of the Sindbis glycoproteins, and galactosylation of vesicular stomatitis virus and cellular glycoproteins occurred to similar respective extents in mutant and parent. Electron microscopic examination of Sindbis-infected DTG 1-5-4 showed a remarkable accumulation of nucleocapsids bound to cisternae adjacent to the Golgi apparatus; virions were observed in the lumina of some of these cisternae. That the alterations in both endocytosis and Golgi-associated steps of viral maturation result from a single genetic lesion indicates that these processes are dependent on a common biochemical mechanism. We suggest that endocytic and secretory pathways may share a common component involved in ion transport.

Animals↗

Does the sympathetic nervous system influence sinus arrhythmia in man? Evidence from combined autonomic blockade.

The influences of vagal and sympathetic efferent activity on sinus arrhythmia in man have been studied in healthy subjects by administration of hyoscine butylbromide and atenolol alone and combined using a microcomputer-linked electrocardiogram (e.c.g.) system. Sinus arrhythmia was quantitated as the S.D. of the R-R interval. Sinus arrhythmia was reduced by hyoscine butylbromide, in some subjects to near abolition, but this end-point was unchanged by pre-treatment with atenolol. Atenolol alone prolonged the mean R-R interval and increased sinus arrhythmia. It is suggested that sinus arrhythmia in man is mediated through vagal efferents alone but that atenolol increases the arrhythmia through a central vagotonic effect.

Adult↗

Effects of secretory stimulation on the Golgi apparatus and GERL of rat parotid acinar cells.

The structure and cytochemistry of the Golgi apparatus and GERL of rat parotid acinar cells was studied after in vivo secretory stimulation with isoproterenol. Discharge of mature secretory granules was complete within 1 hr after isoproterenol injection, but immature granules in the Golgi region or near the lumen were not released. At early times (1-5 hr) after isoproterenol, acid phosphatase (AcPase) activity was markedly increased in GERL and immature secretory granules compared to uninjected controls. GERL appeared increased in extent and numerous continuities with immature granules were observed. Reaccumulation of mature secretory granules was first evident at 5 hr, and was almost complete by 16 hr after isoproterenol. Thiamine pyrophosphatase (TPPase) activity, normally restricted to the trans Golgi saccules, was frequently present in immature granules during this time. Narrow cisternae resembling GERL, occasionally in continuity with immature granules, also contained TPPase reaction product. By 16-24 hr after stimulation, the activity and distribution of AcPase and TPPase were similar to control cells. These results demonstrate the dynamic nature of the Golgi apparatus and GERL in parotid acinar cells, and emphasize the close structural and functional relationship between these two structures.

Acid Phosphatase↗

The Golgi apparatus and GERL during postnatal differentiation of rat parotid acinar cells: an electron microscopic cytochemical study.

Morphological and cytochemical changes in the Golgi apparatus and GERL of differentiating parotid acinar cells were examined in Sprague-Dawley rats from 5 days to young adult. At day 5, the Golgi apparatus consisted of 3-6 narrow saccules, with short segments of GERL lying adjacent to the trans Golgi saccule. As the glands matured, the Golgi apparatus increased in size and the saccules became broadened and fenestrated reaching a maximum from days 15-20. The saccules subsequently narrowed slightly and by day 25 resembled those seen in young adults. Numerous cisternae of GERL could be seen at the trans face during this period. While the glands were maturing, marked changes occurred in the distribution of thiamine pyrophosphatase (TPPase) activity in the Golgi saccules. In the immature cells, TPPase activity was restricted to 1 or 2 trans Golgi saccules. However, by day 10 TPPase could also be localized in immature secretory granules and in GERL-like cisternae. Unreactive segments of GERL were also present. This pattern of localization persisted until day 20, after which the TPPase activity in the GERL-like cisternae diminished gradually until by day 40 TPPase again was localized in 1-2 trans Golgi saccules and an occasional immature secretory granule. Acid phosphatase (AcPase) activity was localized primarily in lysosomes in the very young animals and increased in GERL with age up to day 15. From days 15 to 20 there was a decrease in the amount of activity seen in GERL, but from day 20 on, the AcPase activity increased until it reached that seen in young adult animals. These results indicate that the presence of TPPase activity in GERL-like cisternae and immature secretory granules may be dependent upon the developmental as well as the physiologic state of the acinar cells and lend further support to the suggestion that GERL is derived from the trans Golgi saccules.

Acid Phosphatase↗

[Effects of adrenal medullectomy and passive immunization with an anti-CRF antiserum on the corticotropin secretion in response to stress in Long-Evans rats].

Ether stress-induced ACTH and corticosterone secretion is similar in 60 days adrenal medullectomized and intact Long-Evans Rats. Passive immunisation with a 41-CRF antiserum impairs corticotropic function after stress. This decrease is more pronounced in adrenal medullectomized than in intact animals. These results suggest that epinephrine plays a minor role on the stress induced ACTH secretion. This role can be demonstrated only after immunoneutralisation of endogenous 41-CRF.

Adrenal Medulla↗

Adrenal medullary opiate receptors. Pharmacological characterization in bovine adrenal medulla and a human pheochromocytoma.

We have characterized the opiate binding sites on the membranes of bovine adrenal medulla and human pheochromocytoma, using 3H-labeled D-Ala2-D-Leu5-enkephalin ( [3H]DADLE), [3H]etorphine, and [3H]ethylketocyclazocine ( [3H]EKC). Binding was stereoselective in both membrane preparations. Association and dissociation kinetics showed that steady state was achieved after 20-25 min of incubation at 37 degrees. Saturation experiments were performed in the absence or in the presence of morphiceptin (1 microM), which masks the mu sites, D-Ser2-Leu-enkephalin-Thr6 (100 nM), which masks delta sites, or DADLE (5 microM), which was found to mask the delta, mu, and benzomorphan receptor. Taking into consideration the affinities of the three radioligands used (DADLE identifying the delta and mu sites when used in the nanomolar range; etorphine identifying the delta, mu, and benzomorphan sites; EKC identifying the delta, mu, kappa, and benzomorphan receptors) we have characterized pharmacologically the opiate sites present on bovine and human membranes. Human pheochromocytoma membranes contained (a) mu binding sites (15 fmoles/mg of protein, KD [3H]etorphine 1.0 nM, [3H]EKC 5.4 nM, [3H]DADLE 5.6 nM); (b) kappa sites (41 fmoles/mg of protein, KD [3H]EKC 1.0 nM); (c) benzomorphan sites (115 fmoles/mg of protein, KD [3H]etorphine and [3H]EKC 1.0 nM). On bovine membranes we have detected (a) delta binding sites (10 fmoles/mg of protein, KD [3H]DADLE 0.7 nM); (b) mu sites (24 fmoles/mg of protein, KD [3H]DADLE 2.9 nM, [3H]etorphine 0.2 nM, [3H]EKC 3.4 nM); (c) kappa sites (12 fmoles/mg of protein, KD [3H]EKC 0.4 nM); (d) benzomorphan sites (80 fmoles/mg of protein, KD [3H]etorphine 0.2 nM, [3H]EKC 1.3 nM); (e) a residual high-affinity (20 fmoles/mg of protein, KD 0.2 nM) site identified by [3H]etorphine in the presence of 5 microM DADLE. The relative proportions of benzomorphan sites were equal in both tissues (65% of the high-affinity sites) whereas kappa receptors were more abundant on human membranes (25%) than on bovine membranes (9% of the high-affinity sites).

Adrenal Gland Neoplasms↗

In vitro study of frog (Rana ridibunda Pallas) neurointermediate lobe secretion by use of a simplified perifusion system. III. Effect of neuropeptides on alpha-MSH secretion.

It has been previously demonstrated that thyrotropin-releasing hormone (TRH) stimulates in vitro the release of alpha-melanocyte-stimulating hormone (alpha-MSH) in frog. In the present study, the effects of various neuropeptides on spontaneous and/or TRH-induced alpha-MSH secretion were investigated, using a well-defined perifusion system technique. Vasoactive intestinal peptide, (VIP) a neurohormone which stimulates TRH target cells in mammals, was totally devoid of effect on frog melanotrophs although VIP-like material could be detected in neurointermediate lobe extracts. Somatostatin-like immunoreactive material was found in high concentrations in the frog neurointermediate lobe complex, but synthetic somatostatin (from 10(-10) to 10(-6) M) did not modify the spontaneous release of alpha-MSH. At doses of 10(-8) and 10(-6) M, synthetic somatostatin did not modify TRH-induced alpha-MSH secretion. Morphine (10(-5) M) and opioid peptides (10(-10) to 10(-6) M) had no effect on spontaneous alpha-MSH secretion. In addition, methionine enkephalin (10(-5) M) did not modify the stimulatory effect of TRH on alpha-MSH secretion. From these results we conclude that, among the neuropeptides which modulate prolactin secretion in mammals, only TRH is involved in alpha-MSH secretion in the frog.

Animals↗

Opiate binding sites spectrum on bovine adrenal medullas and six human pheochromocytomas.

Opiate binding sites have been characterized on membranes from bovine adrenal medullas and six human pheochromocytomas. In human tumors, large variations in site distribution were observed. Kappa and benzomorphan sites represented the majority of the sites detected. The heterogeneity of the opiate sites on these tissues could explain the observed differences in the pharmacological responses to opiates of cultured cells from these tissues. Furthermore, adrenal medullas could be a good model for the study of the kappa site action at the cellular level.

Adrenal Gland Neoplasms↗